miR-126-3p contributes to sorafenib resistance in hepatocellular carcinoma via downregulating SPRED1

Wenliang Tan; Zhirong Lin; Xianqing Chen; Wenxin Li; Sicong Zhu; Yingcheng Wei; Liyun Huo; Yajin Chen; Changzhen Shang

doi:10.21037/atm-20-2081

miR-126-3p contributes to sorafenib resistance in hepatocellular carcinoma via downregulating SPRED1

Ann Transl Med. 2021 Jan;9(1):38. doi: 10.21037/atm-20-2081.

Authors

Wenliang Tan^{1

2}, Zhirong Lin^{1

2}, Xianqing Chen³, Wenxin Li⁴, Sicong Zhu^{1

5}, Yingcheng Wei^{1

2}, Liyun Huo^{1

2}, Yajin Chen^{1

2}, Changzhen Shang^{1

2}

Affiliations

¹ Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.
² Department of Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
³ Department of Hepatobiliary Surgery, the Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen, China.
⁴ Department of Cardiology, the Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen, China.
⁵ Department of Surgical Intensive Care Unit, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

Abstract

Background: Sorafenib can prolong the survival of patients with advanced hepatocellular carcinoma (HCC). However, drug resistance remains the main obstacle to improving its efficiency. This study aimed to explore the likely molecular mechanism of sorafenib resistance.

Methods: Differentially expressed microRNAs (miRNAs) related to sorafenib response were analyzed with the Limma package in R software. The expression levels of miR-126-3p and sprouty-related EVH1 domain-containing protein 1 (SPRED1) in HCC cells were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell viability and proliferation were detected with Cell Counting Kit-8 (CCK-8), EdU proliferation, and clone formation assays. Transwell assays were performed to measure cell migration and invasion. TargetScan, MicroRNA Target Prediction Database (miRDB), and StarBase v2.0 were used to predict the targets of miR-126-3p. SPRED1 was confirmed as a target gene of miR-126-3p by dual-luciferase reporter assay and Western blotting. Finally, the in vivo anti-tumor effect of LV-miR-126-3p inhibitor combined with sorafenib was evaluated via subcutaneous tumor models.

Results: HCC cells with high expression of miR-126-3p exhibited increased resistance to sorafenib. The results of bioinformatics analysis and the dual-luciferase reporter assay showed that miR-126-3p directly targeted SPRED1. The sensitivity of HCC cells to sorafenib was markedly enhanced by SPRED1 upregulation. Gain- and loss-of function experiments verified that miR-126-3p induced sorafenib resistance in HCC through downregulating SPRED1. Furthermore, the inhibition of miR-126-3p markedly increased the effectiveness of sorafenib against HCC in vivo. Mechanistically, our results suggested that miR-126-3p promoted sorafenib resistance via targeting SPRED1 and activating the ERK signaling pathway.

Conclusions: Our study demonstrates that regulating the miR-126-3p/SPRED1 axis might be a promising strategy for enhancing the antitumor effect of sorafenib in the treatment of HCC.

Keywords: ERK signaling pathway; hepatocellular carcinoma (HCC); miR-126-3p; sorafenib resistance; sprouty-related EVH1 domain-containing protein 1 (SPRED1).